GSM7178572: SampleZ, Spatial transcriptome sequencing; Homo sapiens; RNA-Seq
Before sequencing, we assessed RNA quality using DV200, DV200≥50% be used. We then conducted a tissue adhesion test, followed by deparaffinization and H&E staining. After staining, we captured images of the area using brightfield settings and released the RNA through decrosslinking. We then added an integrative transcriptome probe panel, which consisted of a pair of specific probes for each target gene, to the previously stained tissue. Probe pairs hybridized to their complementary target RNA and formed a stable ligation product with the addition of a ligase. After processing the ligation structure, we released the single-stranded ligation products from the tissue. RT Master Mix containing reverse transcription reagents was then added for amplification, followed by the addition of UMI, barcode and partial Read1. The library with barcoded ligation products was released to qPCR to determine cycle amplification number, added with index, and finally constructed the library containing P5 and P7 primers used for Illumina amplification. The library molecules were cleaned up by SPRIselect and assessed, quantified, and sequenced on a bioanalyzer or similar instrument using PE150 mode sequencing on the Illumina NovaSeq600 platform Spatial transcriptome sequencing